Altered neurite morphology and cholinergic function of induced pluripotent stem cell-derived neurons from a patient with Kleefstra syndrome and autism

Abstract

The aim of the present study was to establish an in vitro Kleefstra syndrome (KS) disease model using the human induced pluripotent stem cell (hiPSC) technology. Previously, an autism spectrum disorder (ASD) patient with Kleefstra syndrome (KS-ASD) carrying a deleterious premature termination codon mutation in the EHMT1 gene was identified. Patient specific hiPSCs generated from peripheral blood mononuclear cells of the KS-ASD patient were differentiated into post-mitotic cortical neurons. Lower levels of EHMT1 mRNA as well as protein expression were confirmed in these cells. Morphological analysis on neuronal cells differentiated from the KS-ASD patient-derived hiPSC clones showed significantly shorter neurites and reduced arborization compared to cells generated from healthy controls. Moreover, density of dendritic protrusions of neuronal cells derived from KS-ASD hiPSCs was lower than that of control cells. Synaptic connections and spontaneous neuronal activity measured by live cell calcium imaging could be detected after 5 weeks of differentiation, when KS-ASD cells exhibited higher sensitivity of calcium responses to acetylcholine stimulation indicating a lower nicotinic cholinergic tone at baseline condition in KS-ASD cells. In addition, gene expression profiling of differentiated neuronal cells from the KS-ASD patient revealed higher expression of proliferation-related genes and lower mRNA levels of genes involved in neuronal maturation and migration. Our data demonstrate anomalous neuronal morphology, functional activity and gene expression in KS-ASD patient-specific hiPSC-derived neuronal cultures, which offers an in vitro system that contributes to a better understanding of KS and potentially other neurodevelopmental disorders including ASD.

Investigation of de novo mutations in a schizophrenia case-parent trio by induced pluripotent stem cell based in vitro disease-modeling: Convergence of schizophrenia and autism-related cellular phenotypes

“…Both accelerated and decreased neurite outgrowth have also been implicated as important neurodevelopmental cellautonomous phenotypes in ASD and SZ. In an iPSC-based model of Kleefstra-syndrome, a syndromic and genetically well described subtype of ASD, NPCs demonstrated increased proliferation, while neurite arborization was reduced in mature neurons (45). It has also been shown that KHSRP regulates the mRNA-stability of GAP43, an important player in the process of axonal and dendritic growth (46).…”

Identification of De Novo JAK2 and MAPK7 Mutations Related to Autism Spectrum Disorder Using Whole-Exome Sequencing in a Chinese Child and Adolescent Trio-Based Sample

“…Many genes in this pathway are involved in the pathogenesis of many psychiatric disorders, such as ASD (Wen et al 2016), attention deficit and hyperactivity disorder (ADHD), bipolar disorder, and schizophrenia (Zhao et al 2018). Moreover, the protein encoded by MAPK7 is a member of the MAPK family involved in a wide variety of cellular processes, such as proliferation, differentiation, transcription regulation, and brain development (Pearson et al 2001), and abnormalities in these processes can lead to the development of ASD symptoms (Nagy et al 2017;Courchesne et al 2019). Previous evidence also indicated rare mutations in the MAPK gene family participating in ASD, such as MAPK3 (Park et al 2017) and MAPK1 knockout mice showing ASD-like behavior (Satoh et al 2011).…”

Generation of a Urine-Derived Ips Cell Line from a Patient with a Ventricular Septal Defect and Heart Failure and the Robust Differentiation of These Cells to Cardiomyocytes via Small Molecules

“…The method of disease modeling with patient-specific iPSCs can be applied to all types of chromosomal inherited diseases, from monogenetic to complex polygenetic disorders [25][26][27][28]. iPSCs hold great promise for disease modeling and regenerative therapies.…”